Li Zhi-Qin, Gu Xin-Yu, Hu Jin-Xing, Ping Yu, Li Hua, Yan Jing-Ya, Li Juan, Sun Ran, Yu Zu-Jing, Zhang Yi
Department of Infectious Disease, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
Biotherapy Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China School of Life Sciences, Zhengzhou University, Zhengzhou 450052, Henan, China Key Laboratory of Clinical-Medicine, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China
Biosci Rep. 2016 May 20;36(3). doi: 10.1042/BSR20160088. Print 2016 Jul.
It has been suggested that Hepatitis C virus (HCV) core protein is associated with metabolic disorders of liver cell. However, the precise mechanism is still unclear. The aim of the present study was to explore the impact of HCV core protein on hepatocyte metabolism by HepG2 and the possible involvement of long non-coding (lnc) RNAs in this process. The effect of HCV core protein on lncRNAs expression was examined with quantitative RT-PCR (qRT-PCR). Manipulation of HVC core protein and lncRNA HOTAIR was to evaluate the role of interaction between them on cell metabolism-related gene expression and cellular metabolism. The potential downstream Sirt1 signal was examined by western blotting and qRT-PCR. Our data suggested that suppression of HOTAIR abrogates HCV core protein-induced reduction in Sirt1 and differential expression of glucose- and lipid-metabolism-related genes. Also it benefits for metabolic homoeostasis of hepatocyte indicated by restoration of cellular reactive oxygen species (ROS) level and NAD/NADH ratio. By manipulation of HOTAIR, we concluded that HOTAIR negatively regulates Sirt1 expression through affecting its promotor methylation. Moreover, overexpression of Sirt1 reverses pcDNA-HOTAIR-induced glucose- and lipid-metabolism-related gene expression. Our study suggests that HCV core protein causes dysfunction of glucose and lipid metabolism in liver cells through HOTAIR-Sirt1 signalling pathway.
有人提出丙型肝炎病毒（HCV）核心蛋白与肝细胞的代谢紊乱有关。然而，确切机制仍不清楚。本研究的目的是通过HepG2细胞探讨HCV核心蛋白对肝细胞代谢的影响以及长链非编码（lnc）RNA在这一过程中可能的参与情况。采用定量逆转录聚合酶链反应（qRT-PCR）检测HCV核心蛋白对lncRNAs表达的影响。对HVC核心蛋白和lncRNA HOTAIR进行操作，以评估它们之间的相互作用对细胞代谢相关基因表达和细胞代谢的作用。通过蛋白质免疫印迹法和qRT-PCR检测潜在的下游Sirt1信号。我们的数据表明，抑制HOTAIR可消除HCV核心蛋白诱导的Sirt1降低以及葡萄糖和脂质代谢相关基因的差异表达。这也有利于肝细胞代谢稳态，表现为细胞活性氧（ROS）水平和NAD/NADH比值的恢复。通过对HOTAIR的操作，我们得出结论，HOTAIR通过影响其启动子甲基化负向调节Sirt1表达。此外，Sirt1的过表达可逆转pcDNA-HOTAIR诱导的葡萄糖和脂质代谢相关基因的表达。我们的研究表明，HCV核心蛋白通过HOTAIR-Sirt1信号通路导致肝细胞葡萄糖和脂质代谢功能障碍。
